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Tissue print hybridisation and reverse transcriptase PCR in the detection of infectious bursal disease viruses in bursal tissues

Authors
Journal
Research in Veterinary Science
0034-5288
Publisher
Elsevier
Publication Date
Volume
68
Issue
1
Identifiers
DOI: 10.1053/rvsc.1999.0333
Disciplines
  • Medicine

Abstract

Abstract The genome segments of infectious bursal disease viruses ( IBDV ) in the bursa of Fabricius from experimently infected chickens or field samples were detected by tissue print hybridization ( TPH ) with subsequent reverse transcriptase ( RT )- polymerase chain reaction ( PCR ). Bursae were imprinted onto nylon membrane and then hybridized with a cloned digoxigenin ( DIG )-labeled cDNA probe. Tissue prints on nylon membrane were readily distinguished from control prints by color development and differences in signal intensity. In order to verify the TPH test, RT - PCR was used to amplify a 643-base pair fragment on the VP 2 gene of IBDV in the bursa of Fabricius. With all isolates, a c DNA fragment of 643 bp long was generated as expected and further confirmed the specificity of TPH . Our results suggest that a large number of field samples or selected tissues can be rapidly examined by TPH technique when combined with a cloned DIG -labeled c DNA probe.

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